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Evaluation of Different DNA Vaccines against Porcine Reproductive and Respiratory Syndrome (PRRS) in Pigs.

Petrini S, Ramadori G, Villa R, Borghetti P, de Angelis E, Cantoni AM, Corradi A, Amici A, Ferrari M - Vaccines (Basel) (2013)

Bottom Line: Only vaccines A and B were able to reduce the clinical signs of the infection.Neutralizing antibody were detected Post Challenge Days 61 (PCD) in all groups.In the other groups, the IFN-g were detected after challenge infection.

View Article: PubMed Central - PubMed

Affiliation: Umbria and Marche Experimental Zooprophylaxis Institute, via Gaetano Salvemini 1, Perugia 06126, Italy. s.petrini@izsum.it.

ABSTRACT
In veterinary medicine, there have been different experiences with the plasmid DNA vaccination. In this area and with the hypothesis to demonstrate the effectiveness of different plasmids encoding porcine respiratory and reproductive syndrome (PRRS), five DNA vaccines against PRRS were evaluated for their innocuity and efficacy in pigs. Eighteen animals were divided into five groups which were injected with five (A, B, C, D, E) different DNA vaccines. Albeit, none of the proposed vaccines were able to protect the animals against PRRS virus. Only vaccines A and B were able to reduce the clinical signs of the infection. ELISA IgM were detected 30 days after the first vaccination in the pigs injected by Vaccine A or B. ELISA IgG were detected 90 days after the first vaccination in the pigs injected by Vaccine B or C. Neutralizing antibody were detected Post Challenge Days 61 (PCD) in all groups. In the pigs inoculated with Vaccine C, IFN-g were detected 90 days after first vaccination, and after challenge exposure they increased. In the other groups, the IFN-g were detected after challenge infection. Pigs injected with each of the vaccines A, B, C, D and E showed a significantly higher level of CD4(-)CD8⁺ lymphocytes (p < 0.001) after infection in comparison with their controls.

No MeSH data available.


Related in: MedlinePlus

FITC immunofluorescence of mouse embryonic fibroblast cells (NIH-3T3) transfected with pVAX1-48CpG-NeuL-ORF4-Myc. (A) or pVAX1-48CpG-NeuL-ORF5-Myc; (B) after 48 h from transfection.
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vaccines-01-00463-f007: FITC immunofluorescence of mouse embryonic fibroblast cells (NIH-3T3) transfected with pVAX1-48CpG-NeuL-ORF4-Myc. (A) or pVAX1-48CpG-NeuL-ORF5-Myc; (B) after 48 h from transfection.

Mentions: We cloned into the restriction site Xba I of pVAX1-48CpG-neuL-ORF4 and pVAX1-48CpG-neuL-ORF5 a sequence encoding for the antigenic Myc tag epitope EQKLISEEDL. This modification lead to the expression of ORF4 or ORF5 in fusion with the antigen Myc tag that is recognized by a commercial antibody FITC conjugated F2047 (Sigma-Aldrich, Milan, Italy) and then allowed us to follow the expression of ORF4 and ORF5 in mouse embryonic fibroblast cells (NIH-3T3) by confocal microscopy. Transfections of NIH-3T3 using a lipofectamine established protocol (Invitrogen, San Diego, USA) of either pVAX1-48CpG-NeuL-ORF4-Myc or pVAX1-48CpG-NeuL-ORF5-Myc led to a marked cytosolic expression of the encoded antigens (Figure 7A,B).


Evaluation of Different DNA Vaccines against Porcine Reproductive and Respiratory Syndrome (PRRS) in Pigs.

Petrini S, Ramadori G, Villa R, Borghetti P, de Angelis E, Cantoni AM, Corradi A, Amici A, Ferrari M - Vaccines (Basel) (2013)

FITC immunofluorescence of mouse embryonic fibroblast cells (NIH-3T3) transfected with pVAX1-48CpG-NeuL-ORF4-Myc. (A) or pVAX1-48CpG-NeuL-ORF5-Myc; (B) after 48 h from transfection.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4494207&req=5

vaccines-01-00463-f007: FITC immunofluorescence of mouse embryonic fibroblast cells (NIH-3T3) transfected with pVAX1-48CpG-NeuL-ORF4-Myc. (A) or pVAX1-48CpG-NeuL-ORF5-Myc; (B) after 48 h from transfection.
Mentions: We cloned into the restriction site Xba I of pVAX1-48CpG-neuL-ORF4 and pVAX1-48CpG-neuL-ORF5 a sequence encoding for the antigenic Myc tag epitope EQKLISEEDL. This modification lead to the expression of ORF4 or ORF5 in fusion with the antigen Myc tag that is recognized by a commercial antibody FITC conjugated F2047 (Sigma-Aldrich, Milan, Italy) and then allowed us to follow the expression of ORF4 and ORF5 in mouse embryonic fibroblast cells (NIH-3T3) by confocal microscopy. Transfections of NIH-3T3 using a lipofectamine established protocol (Invitrogen, San Diego, USA) of either pVAX1-48CpG-NeuL-ORF4-Myc or pVAX1-48CpG-NeuL-ORF5-Myc led to a marked cytosolic expression of the encoded antigens (Figure 7A,B).

Bottom Line: Only vaccines A and B were able to reduce the clinical signs of the infection.Neutralizing antibody were detected Post Challenge Days 61 (PCD) in all groups.In the other groups, the IFN-g were detected after challenge infection.

View Article: PubMed Central - PubMed

Affiliation: Umbria and Marche Experimental Zooprophylaxis Institute, via Gaetano Salvemini 1, Perugia 06126, Italy. s.petrini@izsum.it.

ABSTRACT
In veterinary medicine, there have been different experiences with the plasmid DNA vaccination. In this area and with the hypothesis to demonstrate the effectiveness of different plasmids encoding porcine respiratory and reproductive syndrome (PRRS), five DNA vaccines against PRRS were evaluated for their innocuity and efficacy in pigs. Eighteen animals were divided into five groups which were injected with five (A, B, C, D, E) different DNA vaccines. Albeit, none of the proposed vaccines were able to protect the animals against PRRS virus. Only vaccines A and B were able to reduce the clinical signs of the infection. ELISA IgM were detected 30 days after the first vaccination in the pigs injected by Vaccine A or B. ELISA IgG were detected 90 days after the first vaccination in the pigs injected by Vaccine B or C. Neutralizing antibody were detected Post Challenge Days 61 (PCD) in all groups. In the pigs inoculated with Vaccine C, IFN-g were detected 90 days after first vaccination, and after challenge exposure they increased. In the other groups, the IFN-g were detected after challenge infection. Pigs injected with each of the vaccines A, B, C, D and E showed a significantly higher level of CD4(-)CD8⁺ lymphocytes (p < 0.001) after infection in comparison with their controls.

No MeSH data available.


Related in: MedlinePlus